Journal: Circulation

Article Title: Smooth Muscle Notch1 Mediates Neointimal Formation After Vascular Injury

doi: 10.1161/circulationaha.108.790485

Figure Lengend Snippet: Figure 1. Expression of Notch receptors in ECs and SMCs. A, Western blot analysis shows the expression of total Notch1 in ECs isolated from hearts of WT, Notch1flox/, smCre-Tg, N1/-, and smN1/ mice. B, Western blot analysis shows the expression of total Notch1 and Notch3 in SMCs isolated from aortas of WT, Notch1flox/, smCre-Tg, N1/, and smN1/ mice. C, Immunofluores- cence staining of Notch1, Notch2, and Notch3 in uninjured carotid artery (left) and ligated injured carotid artery (14 days after ligation; right) of WT mice. Vessels were costained for smooth muscle actin. Smooth muscle actin is stained red, Notch receptors were stained green, and nuclei were stained blue (DAPI). Yellow indicates presence of both red and green staining. White bar20 m. D, Immunofluorescence costaining of smooth muscle actin with activated Notch1 (NICD). Red was SMA, green was NICD, and blue was DAPI. White bar20 m.

Article Snippet: Immunofluorescence for smooth muscle actin (Sigma; A2547), Notch1 (Santa Cruz Biotechnology; sc-6014), activated Notch1 (NICD; Abcam, ab8925), Notch2 (Rockland; 100-401-406), Notch3 (Santa Cruz Biotechnology; sc-7424), cleaved caspase-3 (Cell Signaling Technology; 9664), and proliferating cell nuclear antigen (PCNA) (Laboratory Vision; RB-9055) was performed by standard procedures.

Techniques: Expressing, Western Blot, Isolation, Staining, Ligation

Journal: Journal of Clinical Laboratory Analysis

Article Title: High‐dimensional single‐cell proteomics analysis reveals the landscape of immune cells and stem‐like cells in renal tumors

doi: 10.1002/jcla.23155

Figure Lengend Snippet: Purified antibodies about the stem‐like cell‐centric panel

Article Snippet: Notch2 , 165Ho , MHN225 , Fluidigm , 3165026B.

Techniques: Purification

Journal: Frontiers in Immunology

Article Title: Endothelial cells regulate mesangial cells through the Dll4/Notch3 axis to participate in glomerular injury in lupus nephritis

doi: 10.3389/fimmu.2026.1720756

Figure Lengend Snippet: EC regulate MC via the Dll4/Notch3 axis. (A) ELISA assays were performed to detect the expression of Dll4 in ECs after LPS stimulation. (B) CCK8 assay showing the effects of Dll4 on MC viability. (C) qRT-PCR assay showing Notch3 relative genes were upregulated in MC after Dll4 stimulation. (D) CCK8 assay showing the effects of Tarextumab on MC viability after Dll4 interference. (E) Western blotting analysis of the expression of the Notch3-ICD and its downstream-related proteins HEY1 and HEY2 following interference with Dll4 alone or in combination with Tarextumab. (F) Quantification of Western blotting. Data were representative of three independent experiments (n = 3 per group). (G) Representative images of wound healing assays showing the migration of MC cells treated with Dll4 or Dll4 in combination with Tarextumab. Scale bar = 200 μm. (H) Quantitative comparison of MC cell migration rates among Control, Dll4, and Dll4+Tarextumab treatment groups (n = 3). (I) Transwell analysis of the effect of Dll4 and Tarextumab on MC cell migration. Scale bar = 2 mm for overview. Scale bar = 300 μm for magnified view. (J) Quantitative analysis of the number of migrated cells. Data are presented as mean ± SD (n = 3). *p < 0.05, **p < 0.01. (K) EdU assays were performed to detect the proliferation of MC treated with Dll4 and Tarextumab. Scale bar = 200 μm. (L) Quantitative analysis of MC cell proliferation by EdU assay. Data are presented as mean ± SD (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: After complete attachment, Dll4 (recombinant human Dll4, 100 ng/ml, HEK293 FC, MCE, USA) and Tarextumab (humanized anti-Notch2/3 antibody, 10 ug/ml, OMP 59R5, MCE, USA) were added to MCs.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, CCK-8 Assay, Quantitative RT-PCR, Western Blot, Migration, Comparison, Control, EdU Assay

Journal: Frontiers in Immunology

Article Title: Endothelial cells regulate mesangial cells through the Dll4/Notch3 axis to participate in glomerular injury in lupus nephritis

doi: 10.3389/fimmu.2026.1720756

Figure Lengend Snippet: Dll4 promotes the proliferation and migration of MC via Dll4 in vitro . (A) Co-culture illustration of ECs and MCs according to different treatment, the control group was treated with LPS to ECs alone, siRNA-Dll4 group was siRNA-Dll4 in ECs then treated with LPS, Tarextumab group was ECs treated with LPS and MCs treated with Tarextumab. (B) qRT-PCR results of the Dll4 mRNA expression after Dll4 siRNA transfection in ECs (n = 3, *P < 0.05, **P < 0.01). (C) Transwell assay showing the migration of MC in different groups, Scale bar = 2 mm (upper), Scale bar = 300 μm (lower). (D) MC migration statistics (n = 3, **P < 0.01, ***P < 0.001). (E) EdU assays were performed to characterize the proliferation of MC when knocking down Dll4 in ECs or adding Tarextumab. Scale bar = 150 μm. (F) MC proliferation statistics n = 3 for each group, **P < 0.01, ***P < 0.001.

Article Snippet: After complete attachment, Dll4 (recombinant human Dll4, 100 ng/ml, HEK293 FC, MCE, USA) and Tarextumab (humanized anti-Notch2/3 antibody, 10 ug/ml, OMP 59R5, MCE, USA) were added to MCs.

Techniques: Migration, In Vitro, Co-Culture Assay, Control, Quantitative RT-PCR, Expressing, Transfection, Transwell Assay

Journal: Frontiers in Pharmacology

Article Title: Antiproliferative and Immunoregulatory Effects of Azelaic Acid Against Acute Myeloid Leukemia via the Activation of Notch Signaling Pathway

doi: 10.3389/fphar.2019.01396

Figure Lengend Snippet: Notch was found up-regulated after azelaic acid (AZA) treatment. (A) 120 differential expressed proteins associated in 52 functions after AZA treatment by protein mass spectrometry analysis in the heat map. (B) GO annotation with 528 differential expressed proteins. (C, D) THP-1, Molm-13, NK, and T cells were treated with 10 µM AZA for 24 h, the expression level of ICN1 and ICN2 was validated by western blot. (E) The expression of Notch1 and Notch2 in mouse spleen measured by ICH. IHC, immunohistochemistry.

Article Snippet: The following antibodies were used: CD3(Cat# 100203), CD4 (Cat# 100431), CD8 (Cat# 100761), CD107a (Cat# 328605), TRAIL (Cat# 308205), CD25 (Cat# 302605), and CD69 (Cat# 310903) were purchased from BioLegend (USA), ICN1 (Cat# CSB-PA084572), ICN2 (Cat# CSB-PA964902) were from CUSABIO (USA); GAPDH (Cat# 6004-1), β-actin (Cat# 14395-1) were from ProteinTech (USA).

Techniques: Mass Spectrometry, Expressing, Western Blot, Immunohistochemistry

Journal: Frontiers in Pharmacology

Article Title: Antiproliferative and Immunoregulatory Effects of Azelaic Acid Against Acute Myeloid Leukemia via the Activation of Notch Signaling Pathway

doi: 10.3389/fphar.2019.01396

Figure Lengend Snippet: Azelaic acid (AZA) exerts anti-leukemic effect by activating the Notch signaling pathway. (A) Notch responsive elements were transfected into 293T cells after 24 h. Cells were then treated with 10 µM AZA, 10 µM RO4929097, and combination for 24 h, the Notch signaling reporter assay was measured by dual luciferase reporter activity. (B) Validation of the RNA expression of Notch1 and Notch2, the downstream target genes HES1 and HEY1 in Molm-13 and THP-1 cells by qPCR. (C) The protein expression level of ICN1, ICN2, HEY1, and HES1 in acute myeloid leukemia (AML) cell lines after treatment of AZA and RO4929097 and their detection by western blot. ImageJ was used for the densitometric analysis. Data represent means ± SD. (D) Molm-13 cells were pretreated with 10 µM RO4939097 for 24 h, and then treated with 10 µM AZA for another 24 h. Cells were collected for apoptosis analysis. (E) NK cells and T cells were pretreated with 10 µM AZA, 10 µM RO4929097, and combination for 24h before co-culture with THP-1 cell and Molm-13 cells at an E:T ratio of 5:1. The cytotoxicity of NK and T was determined by detecting the LDH release rate. (F) NK and T cells were pre-treated with AZA and RO4929097, then co-cultured with THP-1 cell at an E:T ratio of 3:1 for 4 h. The level of TNF-α and IFN-γ in the supernatant was measured by ELISA. A Total of three independent experiments were performed. *P < 0.05, **P < 0.01,***P < 0.001.

Article Snippet: The following antibodies were used: CD3(Cat# 100203), CD4 (Cat# 100431), CD8 (Cat# 100761), CD107a (Cat# 328605), TRAIL (Cat# 308205), CD25 (Cat# 302605), and CD69 (Cat# 310903) were purchased from BioLegend (USA), ICN1 (Cat# CSB-PA084572), ICN2 (Cat# CSB-PA964902) were from CUSABIO (USA); GAPDH (Cat# 6004-1), β-actin (Cat# 14395-1) were from ProteinTech (USA).

Techniques: Transfection, Reporter Assay, Luciferase, Activity Assay, Biomarker Discovery, RNA Expression, Expressing, Western Blot, Co-Culture Assay, Cell Culture, Enzyme-linked Immunosorbent Assay